Coupling metal stable isotope compositions and X-ray absorption spectroscopy to study metal pathways in soil-plant systems: a mini review.

Excess and limited trace metal contents in soils and plants can limit crop yields and pose a risk for the environment and human health. This mini-review reports on the emerging approach of combining X-ray absorption spectroscopy (XAS) with isotope analyses to improve the understanding of metal speciation and dynamics in soil-plant systems. In soils and their components, shifts in isotope compositions could be in some cases linked to changing metal speciation and thereby provide information on processes that control the phytoavailability of metals. In plants, the XAS-isotope approach has potential to improve the understanding of how complex interactions of metal speciation, redox processes, and membrane transport control metal uptake and translocation to edible plant parts. Yet, the XAS-isotope approach proves to be in a rather exploratory phase, and many research gaps remain. Such limitations can be overcome by methodological improvements and combining the approach with molecular biology and modelling approaches.

Revealing the pathways of cadmium uptake and translocation in cacao trees (Theobroma cacao L.): A 108Cd pulse-chase experiment.

The pathways through which cadmium (Cd) is taken up and loaded into cacao beans (nibs) are yet to be revealed. Previous work suggested that Cd loading into cacao nibs may occur via direct xylem uptake rather than phloem-mediated redistribution from the leaves. A stable isotope (108Cd) pulse-chase experiment was set up to identify the pathways of Cd loading into cacao nibs. The topsoil beneath two mature cacao trees in the field was enriched in 108Cd via surface irrigation with a spiked solution. The increase in 108Cd isotopic abundance (IA) in the plant tissues was followed up for 548 days after spiking. The 108Cd IA in the plant tissues increased from natural abundance (0.89 %) to 7.0 % (tree A) and 10.1 % (tree B) at equilibrium. The tracer was taken up in the plant tissues in the order immature leaves > mature leaves > nibs in both trees, while tracer uptake in flowers and cherelles was less consistent between the trees. Half of the equilibrium 108Cd IA was reached in the nibs at 191 days after spiking, significantly later than corresponding values for mature (151 days) and immature leaves (117 days). Pod maturation from flower stage takes about 6 months with most Cd entering the nibs at the last stage of development. The rather slow rise in the 108Cd IA in the nibs compared to the leaves hence suggests that Cd in cacao nibs likely originates from phloem-redistribution from the stem, branches or mature leaves and not from direct root-to-nib transport via the xylem.

From soil to cacao bean: Unravelling the pathways of cadmium translocation in a high Cd accumulating cultivar of Theobroma cacao L.

The research on strategies to reduce cadmium (Cd) accumulation in cacao beans is currently limited by a lack of understanding of the Cd transfer pathways within the cacao tree. Here, we elucidated the transfer of Cd from soil to the nib (seed) in a high Cd accumulating cacao cultivar. Here, we elucidated the transfer of Cd from soil to the nib (seed) in a high Cd accumulating cacao cultivar through Cd stable isotope fractionation, speciation (X-Ray Absorption Spectroscopy), and localization (Laser Ablation Inductively Coupled Plasma Mass Spectrometry). The plant Cd concentrations were 10-28 higher than the topsoil Cd concentrations and increased as placenta< nib< testa< pod husk< root< leaf< branch. The retention of Cd in the roots was low. Light Cd isotopes were retained in the roots whilst heavier Cd isotopes were transported to the shoots (Δ 114/110 Cd shoot-root = 0.27 ± 0.02 ‰ (weighted average ± standard deviation)). Leaf Cd isotopes were heavier than Cd in the branches (Δ 114/110 Cd IF3 leaves-branch = 0.18 ± 0.01 ‰), confirming typical trends observed in annual crops. Nibs and branches were statistically not distinguishable (Δ 114/110 Cd nib-branch = -0.08‰ ± 0.06 ‰), contrary to the leaves and nibs (Δ 114/110 Cd nib-IF3 leaves = -0.25‰ ± 0.05 ‰). These isotope fractionation patterns alluded to a more direct transfer from branches to nibs rather than from leaves to nibs. The largest fraction (57%) of total plant Cd was present in the branches where it was primarily bound to carboxyl-ligands (60-100%) and mainly localized in the phloem rays and phelloderm of the bark. Cadmium in the nibs was mainly bound to oxygen ligands (60-90%), with phytate as the most plausible ligand. The weight of evidence suggested that Cd was transferred like other nutrients from root to shoot and accumulated in the phloem rays and phelloderm of the branches to reduce the transfer to foliage. Finally, the data indicated that the main contribution of nib Cd was from the phloem tissues of the branch rather than from leaf remobilization. This study extended the limited knowledge on Cd accumulation in perennial, woody crops and revealed that the Cd pathways in cacao are markedly different than in annual crops.

Changes of Cadmium Storage Forms and Isotope Ratios in Rice During Grain Filling.

Rice poses a major source of the toxic contaminant cadmium (Cd) for humans. Here, we elucidated the role of Cd storage forms (i.e., the chemical Cd speciation) on the dynamics of Cd within rice. In a pot trial, we grew rice on a Cd-contaminated soil in upland conditions and sampled roots and shoots parts at flowering and maturity. Cd concentrations, isotope ratios, Cd speciation (X-ray absorption spectroscopy), and micronutrient concentrations were analyzed. During grain filling, Cd and preferentially light Cd isotopes were strongly retained in roots where the Cd storage form did not change (Cd bound to thiols, Cd-S = 100%). In the same period, no net change of Cd mass occurred in roots and shoots, and the shoots became enriched in heavy isotopes (Δ114/110Cd maturity-flowering = 0.14 ± 0.04‰). These results are consistent with a sequestration of Cd in root vacuoles that includes strong binding of Cd to thiol containing ligands that favor light isotopes, with a small fraction of Cd strongly enriched in heavy isotopes being transferred to shoots during grain filling. The Cd speciation in the shoots changed from predominantly Cd-S (72%) to Cd bound to O ligands (Cd-O, 80%) during grain filling. Cd-O may represent Cd binding to organic acids in vacuoles and/or binding to cell walls in the apoplast. Despite this change of ligands, which was attributed to plant senescence, Cd was largely immobile in the shoots since only 0.77% of Cd in the shoots were transferred into the grains. Thus, both storage forms (Cd-S and Cd-O) contributed to the retention of Cd in the straw. Cd was mainly bound to S in nodes I and grains (Cd-S > 84%), and these organs were strongly enriched in heavy isotopes compared to straw (Δ114/110Cd grains/nodes- straw = 0.66-0.72‰) and flag leaves (Δ114/110Cd grains/nodes-flag leaves = 0.49-0.52‰). Hence, xylem to phloem transfer in the node favors heavy isotopes, and the Cd-S form may persist during the transfer of Cd from node to grain. This study highlights the importance of Cd storage forms during its journey to grain and potentially into the food chain.

Surface soil liming reduces cadmium uptake in cacao seedlings but subsurface uptake is enhanced.

Cadmium concentrations in cacao (Theobroma cacao L.) beans from South America often exceed trade limits. Liming soil is advocated as a remediation option, but amendments cannot be incorporated into the entire root zone without harming the trees. An experiment was set up to identify how Cd uptake varies within the root zone when surface and subsurface soil layers are either limed or not. The experiment used 22-cm-height pots with top and bottom layers using surface and subsurface soil samples from a cacao field. The potted soils were either surface limed or not or fully limed and layers spiked with stable 108 Cd isotope in various combinations to trace the plant Cd provenance. The root distribution was neither affected by liming nor by soil source; 70% of the root biomass was present in the top layer. Plants grown on the fully limed surface soil had 1.7 times lower Cd concentrations in leaves than the unlimed treatments, whereas this concentration was 1.2 times lower when only the top layer was limed (surface soil used in both layers). The isotope dilution data showed that surface soil liming enhanced Cd uptake from the unlimed bottom layer compared with the unlimed soil, suggesting compensating mechanisms. The pots containing surface soil over subsurface soil also showed that compensating effect but, due to lower phytoavailable Cd in the subsurface soil, surface liming still effectively reduced foliar Cd. We conclude that liming might be a feasible mitigation strategy, but its effectiveness is limited when Cd phytoavailability remains untreated in the subsurface layer.

Arsenic contamination in rainwater harvesting tanks around Lake Poopó in Oruro, Bolivia: An unrecognized health risk.

Drinking water sources used by largely rural and indigenous communities around Lake Poopó in the Bolivian Altiplano are impacted by drought and a combination of natural and anthropogenic mining-related contaminants putting the long-term health and sustainability of these communities at risk. As an alternative drinking water source, 18 rainwater harvesting tanks connected to corrugated iron roofs, each with a first-flush system, were installed in 5 communities around the lake. The water quality of these tanks was monitored over 22 months and compared to alternative unprotected surface and groundwater sources the communities previously relied upon. The rainwater quality was found to be within the Bolivian and World Health Organization (WHO) limits, except for elevated arsenic concentrations two times the recommended health limit (0.01 mg/L). Tracing arsenic concentrations through the rainwater flow-path showed that the elevated arsenic concentrations result from mineral dust particles entering the system when rainwater interacts with the roof catchment, with arsenic leaching out. A leaching test showed that 24 h of contact time between 200 mL of water and <1 g of roof dust is enough to raise the arsenic levels of the water above the Bolivian and WHO limit. Currently, no other research exists evaluating the quality of harvested rainwater in the Bolivian Altiplano for human consumption or the source of arsenic in harvested water. This represents a significant knowledge gap for future development practitioners and programs addressing water security around Lake Poopó and the wider region. As a result, it is strongly recommended to include arsenic as a standard parameter in water quality monitoring of rainwater harvesting projects, especially in active mining regions, and to optimize strategies to minimize roof dust from entering the collection system.